Brown marine algae Gongolaria baccata extract protects Caco-2 cells from oxidative stress induced by tert-butyl hydroperoxide

https://doi.org/10.1016/j.fct.2021.112460Get rights and content

Highlights

  • Antioxidant activity of Gongolaria baccata extract in Caco-2 cells.

  • G. baccata extract prevents NQO1 and GST decreased activities and GSH levels induced by tert-BOOH.

  • G. baccata extract prevents ROS and MDA increased levels induced by tert-BOOH.

  • G. baccata extract prevents caspase 3/7 activity induced by tert-BOOH.

  • GSTM2/Nrf2/AKT1 pathways play essential role in the protective mechanism effect of the extract.

Abstract

Gongolaria baccata (S.G. Gmelin) is marine brown seaweed mainly found on the coasts of the Baltic Sea south to the Mediterranean Sea, Canary Islands, Mauritania and Western Sahara. Herein, we report the cell viability and protective effects attributed to molecular mechanisms underlying antioxidant response to survive oxidative stress injuries. Caco-2 cells were submitted to oxidative stress by treatment with tert-butylhydroperoxide (tert-BOOH). The extract prevented cell damage and enhanced activity of antioxidant defenses (NQO1 and GST activities and GSH levels) reduced by treatment with tert-BOOH. The increases of MDA levels, the amount of intracellular ROS and caspase 3/7 activity induced by tert-BOOH were prevented when cells were treated with the G. baccata extract. Moreover, G. baccata extract caused up-regulation of GSTM2, Nrf2, and AKT1 gene expressions, as well as G. baccata extract reduced significantly Bax, BNIP3, APAF1, ERK1, JNK1, MAPK1, P38, P53, NFκB1, TNFα, IL-6, IL-1β and HO-1 gene expressions related to apoptosis, proinflammation and oxidative stress induced by tert-BOOH. These results suggest that G.baccata extract protected the cells against oxidative damage and inflammation; protective effects that could be linked to their bioactive constituents. Hence, this brown seaweed G.baccata extract could be used for the development of functional foods and/or nutraceuticals.

Introduction

A great deal of interest has been developed to isolate novel bioactive compounds from marine resources because of their numerous health beneficial effects. Marine natural products are recognized as powerful reservoirs of novel, chemically diverse molecules with wide applicability to health sciences (Tempone et al., 2011). Cystoseira C. Agardh, 1820 is a polyphyletic genus of marine macro-algae of the Sargassaceae family. Distributed along the Atlantic-Mediterranean coasts, this genus currently encompasses around 40 species (Guiry and Guiry, 2021) with several bioactivities such as antiinflammatory, antiproliferative, antioxidant (Mhadhebi et al., 2011), antifungal (Lopes et al., 2013), antibacterial (Tajbakhsh et al., 2011), antiprotozoal and antimycobacterial (Spavieri et al., 2010). The Cystoseira species have been recently reassigned into two distinct genera, namely Gongolaria and Ericaria. Cystoseira baccata (S.G. Gmelin), previously named Treptacantha baccata (S.G. Gmelin) has been finally renamed as Gongolaria baccata (S. G. Gmelin) (Molinari-Novoa and Guiry, 2020; Orellana et al., 2019). G. baccata occurs from the Baltic Sea south to the Mediterranean Sea, the Canary Islands, Mauritania and Western Sahara.

Seaweed consumption has been associated with a range of health benefits, such as anticancer, immune-modulatory, antiviral, antibacterial and antioxidant activities which are attributed to seaweed constituents, such as sulphated polysaccharides, carrageenans, fucoidans, terpenoids, polyphenols and polyunsaturated fatty acids (PUFAs) (Smit, 2004). The global production of seaweeds is mostly used for human consumption (80%), and about 20% for the hydrocolloid (agar, alginates and carrageenans) production as functional ingredients in commercial applications as stabilizers, emulsifiers, thickening agents and texture modifiers (EUMOFA, 2018). The economic potential of the algae industry is widely recognized and their bioactives have recently gained a considerable interest due to their multiple applications in the food and/or pharmaceutical industries. Chemically, the Gongolaria genus, previously assigned to Cystoseira genus, contains a wide variety of secondary metabolites, such as terpenoids, steroids, phlorotannins, phenolic compounds as well as other chemical components as, for instance, carbohydrates, triacylglycerols/fatty acids, pigments and vitamins (Bruno de Sousa et al., 2017), with radical scavenging activity (Ammar et al., 2015) and antioxidant activity (Balboa et al., 2013; Custódio et al., 2016; Mhadhebi et al., 2011, 2014). The necessity of antioxidants in food and cosmetics is evident and the search for natural alternative sources is increasing. Because of G.baccata extracts reveal to be a promising source of compounds with potential against oxidative stress, the aim of this research study was to establish if extracts of brown seaweeds (G. baccata) protect against tert-BOOH-induced oxidative stress in Caco-2 cells. As indexes of cellular oxidative stress, the generation of reactive oxygen species (ROS), the content of malondialdehyde (MDA), the activity of antioxidant/detoxification enzymes (NQO1, GST, GSH) and messenger RNA (mRNA) gene expressions linked to apoptosis/oxidative mediators were determined. Colon carcinoma (Caco-2) cells have been widely used as an in vitro model for the large intestine and are a well-established model for investigating the antioxidant effect of bioactive compounds (Phelan et al., 2009; Sambuy et al., 2005).

Section snippets

Chemicals and reagents

The compounds tert-butyl hydroperoxide (tert-BOOH), 3-[4,5 dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT), menadione, dicoumarol, malondialdehyde tetrabutylammonium salt (MDA), dimethyl sulfoxide (DMSO), Dulbecco's phosphate buffered saline (DPBS, D8537), fetal bovine serum (FBS), 2-thiobarbituric acid (TBA), sodium dodecyl sulfate (SDS), β-nicotinamide adenine dinucleotide phosphate (NADPH), flavin adenine dinucleotide (FAD), bovine serum albumin (BSA), N-(2-hydroxyethyl)

Results and discussion

There is growing interest in the use of natural products to aid in the maintenance of human health. This research study demonstrates that G. baccata extract has the ability to protect human intestinal cells against oxidative insult by modulating GSH concentration, ROS generation, MDA production, antioxidant enzyme activities and apoptosis, proinflammation and oxidative stress signaling pathways.

Conclusion

In summary, our results support previous data on the antioxidant effect of G. bacatta extract. In the light of our findings, we concluded that G. baccata extract has the ability to protect human intestinal Caco-2 cells against oxidative insult by modulating GSH concentration, ROS generation, MDA production and antioxidant enzyme activities (NQO1, GST). In addition, this research study demonstrates that the GSTM2, Nrf2 and AKT1 pathways play an essential role in the mechanism underlying the

CRediT authorship contribution statement

María-Aránzazu Martínez: Conceptualization, Funding acquisition, Methodology, Validation, Formal analysis, Investigation, Writing – review & editing. Irma Ares: Investigation, Formal analysis, Writing – original draft, Writing – review & editing. Marta Martínez: Investigation, Formal analysis, Writing – original draft, Writing – review & editing. Bernardo Lopez-Torres: Methodology, Investigation. Jorge-Enrique Maximiliano: Methodology, Investigation. Jose-Luis Rodríguez: Methodology,

Declaration of competing interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgments

This work was supported by the Project Ref. RTA2015-00010-C03 from the Ministerio de Economía, Industria y Competitividad, Spain, and Project Ref. PID 2020-15979RR-C33 from the Ministerio de Ciencia e Innovación, Spain.

References (66)

  • M.M. Gaschler et al.

    Lipid peroxidation in cell death

    Biochem. Biophys. Res. Commun.

    (2017)
  • X. Guinda et al.

    Spatial distribution pattern analysis of subtidal macroalgae assemblages by a non-destructive rapid assessment method

    J. Sea Res.

    (2012)
  • P.J. Hissin et al.

    A fluorimetric method for determination of oxidized and reduced glutathione in tissues

    Anal. Biochem.

    (1976)
  • T. Iantomasi et al.

    Glutathione metabolism in Crohn's disease

    Biochem. Med. Metab. Biol.

    (1994)
  • A.K. Jaiswal

    Nrf2 signaling in coordinated activation of antioxidant gene expression

    Free Radic. Biol. Med.

    (2004)
  • J.A. Juanes et al.

    Macroalgae, a suitable indicator of the ecological status of coastal rocky communities in the NE Atlantic

    Ecol. Indicat.

    (2008)
  • O.H. Lowry et al.

    Protein determination with Folin phenol reagent

    J. Biol. Chem.

    (1951)
  • D. Martin et al.

    Regulation of heme oxygenase-1 expression through the phosphatidylinositol 3-kinase/Akt pathway and the Nrf2 transcription factor in response to the antioxidant phytochemical carnosol

    J. Biol. Chem.

    (2004)
  • R. Masella et al.

    Novel mechanisms of natural antioxidant compounds in biological systems: involvement of glutathione and glutathione-related enzymes

    J. Nutr. Biochem.

    (2005)
  • K. Nicholson et al.

    The protein kinase B/Akt signaling pathway in human malignancy

    Cell. Signal.

    (2002)
  • C. Ramakers et al.

    Assumption-free analysis of quantitative real-time polymerase chain reaction (PCR) data

    Neurosci. Lett.

    (2003)
  • V. Valencia et al.

    Hydrography of the southeastern bay of biscay

  • H. Wang et al.

    Quantifying cellular oxidative stress by dichlorofluorescein assay using microplate reader

    Free Radic. Biol. Med.

    (1999)
  • E. Wells et al.

    The use of macroalgal species richness and composition on intertidal rocky seashores in the assessment of ecological quality under the European Water Framework Directive

    Mar. Pollut. Bull.

    (2007)
  • H.H. Ammar et al.

    Physico-chemical characterization and pharmacological evaluation of sulfated polysaccharides from three species of Mediterranean brown algae of the genus Cystoseira

    Daru

    (2015)
  • M. Brentnall et al.

    Caspase-9, caspase-3 and caspase-7 have distinct roles during intrinsic apoptosis

    BMC Cell Biol.

    (2013)
  • J. Brodie et al.

    A revised check-list of the seaweeds of Britain

    J. Mar. Biol. Assoc. U.K.

    (2015)
  • L. Custódio et al.

    Methanol extracts from Cystoseira tamariscifolia and Cystoseira nodicaulis are able to inhibit cholinesterases and protect a human dopaminergic cell line from hydrogen peroxide-induced cytotoxicity

    Pharm. Biol.

    (2016)
  • A.T. Diplock et al.

    Functional food science and defense against reactive oxygen species

    Br. J. Nutr.

    (1998)
  • M. Dubois et al.

    Colorimetric method for determination of sugars and related substances

    Anal. Chem.

    (1956)
  • A.L. Eggler et al.

    Molecular mechanisms of natural products in chemoprevention: induction of cytoprotective enzymes by Nrf2

    Mol. Nutr. Food Res.

    (2008)
  • Blue Bioeconomy. Situation Report and Perspectives

    (2018)
  • R.L. Fletcher

    Seaweeds of the British Isles. Vol 3 Fucophyceae (Phaeophyceae) Part 1

    (1987)
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